Plants are of incredible importance not only as a crucial pillar of nutrients, but also because of the vast environmental roles they play across an entire ecosystem. It should come as no surprise that a deeper understanding of plants is needed to propel agricultural research in many areas. Whether an agricultural biotech company is looking at plant genomics to increase crop sustainability, or scientists are looking to identify genes related to disease resistance, extracting DNA is a fundamental process that unveils the genetic makeup of these vital community members.
The first step of a plant DNA extraction revolves around cell lysis. While a variety of techniques exist to accomplish this, bead beating stands out as one of the most powerful and efficient methods. Utilizing bead media and high force, bead beating disrupts plant cells, liberating their precious DNA for further extraction and analysis. Let’s delve into the fascinating world of bead beating and its role in unraveling the genetic secrets of plants.
First and foremost, it is important to understand the unique challenges that come with preparing plant tissue for genomic DNA extraction. Plant cells are protected by rigid cell walls composed of cellulose, making them more challenging to break open than animal cells. Bead beating overcomes this challenge by providing the necessary mechanical force to break down the cell walls. When the beads are agitated within the plant tissue, they create a high-impact environment, efficiently shattering the cell walls and releasing the cellular contents in a much more efficient and thorough method compared to alternate techniques, such as mortar and pestle.
Bead beating is a mechanical method that involves the use of bead media along with a buffer solution. The combination of beads and buffer acts as a dynamic duo to disrupt the tough cell walls of plant tissues and facilitate the release of DNA. With the wide range of bead beating solutions that OMNI offers, the sample prep possibilities range from handheld units capable of processing a single sample all the way up to robust homogenizers that can tackle multiple 96 well plates for high throughput needs, as well as a wide variety of bead tube consumables to accommodate any sample.
When it comes to plant material, their core composition contains a variety of downstream-inhibiting compounds that can make or break a sample prep method if not removed adequately during sample preparation steps. Thus, it is no secret that selecting the appropriate buffer to use during the bead beating process can further enhance the effectiveness of a given method. Cetyl trimethyl ammonium bromide, or as it is commonly referred to as CTAB, can dramatically improve the homogenization process and increase the nucleic acid yield by helping push forward the lysis process and removing inhibitory contaminants.The appropriate buffer helps create an optimal environment for DNA stability while also assisting in the solubilization and preservation of the DNA during the extraction process.
Once the plant cells have been effectively disrupted, the DNA is released into the buffer solution along with other cellular components. The next step is to extract and purify the DNA from the mixture, separating it from proteins, RNA, and other cellular debris. Much like the lysis stage, there are numerous techniques centered around the extraction process. Automating the extraction contains several benefits with improved consistency and increased throughput being a huge advantage. Once extracted, this purified DNA can then be subjected to further analyses, such as PCR, DNA sequencing, or genetic modification studies.
The effectiveness of a high throughput plant DNA workflow was demonstrated in a recent OMNI application note featuring a variety of staple crops. In the application note, we’ve showcased how bead beating may benefit a customer with high throughput needs. To summarize our methods, seed radicals were sprouted from corn, wheat and rice and processed in OMNI pre-filled 96 well plates containing 1.4 mm ceramic bead media. Samples were homogenized on the Bead Ruptor™ 96 Bead Mill Homogenizer. Once processed, DNA was then extracted utilizing the chemagic™ 360 automated nucleic acid extractor, and resulting samples demonstrated excellent concentration and purity that aligns with requirements for DNA sequencing workflows.
Check out the Application note below to learn more:
High Throughput DNA Extraction from Staple Crops Utilizing the Bead Ruptor™ 96 Bead Mill Homogenizer and chemagic™ 360 Nucleic Acid Extractor
By utilizing the OMNI Bead Ruptor™ 96 Homogenizer for sample preparation, a user can improve efficiency, consistency, and trust that the homogenate is suitable for downstream needs. Bead beating is a powerful tool for preparing plant samples and incorporating this methodology into workflows around the world could dramatically increase our understanding of these crucial species, propelling us forward in the exploration of nature’s genetic code. As we continue to delve into the genetic intricacies of plants, we uncover the keys to their resilience, diversity, and the vital role they play in sustaining life on Earth.
For research use only. Not for use in diagnostic procedures.
